Through phosphorylation-dependent binding to target proteins, 14-3-3 proteins regulate a wide range of physiological processes, including cell proliferation, cell cycle progression, suppression of apoptotic program, and neuronal functions. In order to generate research tools for study the functions of 14-3-3 in various biological systems and potentially to develop 14-3-3-targeted therapeutic strategies, we have developed an AlphaScreen technology based high throughput assay for isolation of small molecules that can disrupt the 14-3-3/client protein interactions. We have optimized and miniaturized the assay for adaptation in a 384-well plate format for robotic operations. The goal of this proposal is to utilize this assay in the Molecular Libraries Screening Centers Network for isolation of small molecule compounds that can interfere with the interaction of 14-3-3 with its client proteins. [unreadable] [unreadable]